刀豆蛋白A诱导人脐静脉内皮细胞表达p-ERK1/2

摘要

Objective To explore the influence of concanavalin A (ConA) on the expression of phosphorylated extra-cellular signal-regulated kinase (p-ERK) 1/2 in CRL2480 cells (human umbilical vein endothelial cell line). Methods Activat-ed effect of ConA on CRL2480 endothelial cells was detected by MTT assay. P-ERK1/2 expression was examined by western blot. The distribution of p-ERK1/2 was observed via immunofluorescent staining. Results After 8h incubation, ConA in-creased the activated index of the cells in a dose dependent manner in the range of 5~20μg/ml (P<0.05). The immu-noblot displayed that 20 μg/ml ConA increased p-ERK1/2 expression in the cells (P<0.05). PD98059 and U0126 (both ERK pathway inhibitors) exerted inhibitory action on p-ERK1/2 expression (P<0.05). The fluo-rescent intensities existed in the cytoplasm and the nuclei of ConA-treated cells were much stronger than those of control cells. Conclusion ConA is able to upregulate p-ERK1/2 expression in CRL2480 cells.%目的：探讨刀豆蛋白A (Concanavalin A，ConA)对CRL2480细胞(人脐静脉内皮细胞系)细胞外调节蛋白激酶(Extracellular signal-regulated kinase，ERK)1/2磷酸化的影响。方法采用MTT法检测ConA对CRL2480细胞活化的影响；采用Western印迹法检测CRL2480细胞磷酸化(p)-ERK1/2的表达；采用细胞免疫荧光技术检测p-ERK1/2在细胞内的分布。结果在5~20μg/ml范围，ConA刺激CRL2480细胞8 h的活化指数具有剂量依赖性；20μg/ml ConA显著增加CRL2480细胞的活化(P<0.05)。ConA能够增加CRL2480细胞的p-ERK1/2表达水平(P<0.05)，ERK途径抑制剂PD98059和U0126明显抑制p-ERK1/2表达。免疫荧光染色结果显示ConA刺激细胞的荧光信号明显高于对照细胞，荧光分布于细胞浆、细胞核周围和细胞核内。结论 ConA能够激活CRL2480细胞表达p-ERK1/2。